Functionalized heterocyclic compounds as antiviral agents

ABSTRACT

The present invention discloses compounds of Formula (I), or pharmaceutically acceptable salts, thereof:which inhibit the protein(s) encoded by hepatitis B virus (HBV) or interfere with the function of the HBV life cycle of the hepatitis B virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HBV infection. The invention also relates to methods of treating an HBV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.62/990,118, which was filed on Mar. 16, 2020. The entire teachings ofthe above application are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceuticalcompositions useful as hepatitis virus replication inhibitors.Specifically, the present invention relates to tetracyclic pyridonecompounds that are useful in treating viral infections such as hepatitisB virus (HBV). The invention provides novel tetracyclic pyridonecompounds as disclosed herein, pharmaceutical compositions containingsuch compounds, and methods of using these compounds and compositions inthe treatment and prevention of HBV infections.

BACKGROUND OF THE INVENTION

Over 240 million people throughout the world are chronically infectedwith hepatitis B virus (HBV). Out of this patient population, at least 2million reside in the United States. For those that are chronicallyinfected, many will develop complications of liver disease fromcirrhosis or hepatocellular carcinoma (HCC).

HBV is a member of the Hepadnavirus family, and it is able to replicatethrough the reverse transcription of an RNA intermediate. The 3.2-kb HBVgenome exists in a circular, partially doublestranded DNA conformation(rcDNA) that has four overlapping open reading frames (ORF). Theseencode for the core, polymerase, envelope, and X proteins of the virus.rcDNA must be converted into covalently closed circular DNA (cccDNA) incells prior to the transcription of viral RNAs. As rcDNA istranscriptionally inert, cccDNA is the only template for HBVtranscription, and its existence is required for infection.

The HBV viral envelope contains a mixture of surface antigen proteins(HBsAg). The HBsAg coat contains three proteins that share a commonregion that includes the smallest of the three proteins (SHBsAg). Theother two proteins, Medium HBsAg (MHBsAg) and Large HBsAg (LHBsAg), bothcontain a segment of SHBsAg with additional polypeptide segments.SHBsAg, MHBsAg, and LHBsAg can also assemble into a non-infectioussubviral particle known as the 22-nm particle that contains the sameproteins found around infectious viral particles. As the 22-nm particlescontain the same antigenic surface proteins that exist around theinfectious HBV virion, they can be used as a vaccine to produceneutralizing antibodies.

In chronically infected patients, the non-infectious 22-nm particles arefound in much greater abundance than the infectious virions. As aresult, the 22-nm particles are thought to be able to protect theinfectious virions from the infected host's immune response. Not onlycan they serve as infectious decoys, but they also suppress normalfunctioning of immune cells thereby impairing the host's immune responseto HBV. Therefore, reducing the level of subviral particles is afeasible therapeutic approach to treating HBV infections. (Refer toWO2015/13990).

In the clinical setting, a diagnostic marker of chronic HBV infection ishigh serum levels of HBsAg. In recent years, data have suggested thatsustained virologic response (SVR) corresponds with HBsAg decline duringearly treatment, while sustained exposure to HBsAg and other viralantigens might lead to inept immunogenicity. Patients that displayhigher decreases in serum HBsAg reached a considerably higher SVRfollowing treatment.

Current treatment options for chronically infected HBV patients arelimited in number and scope. They include interferon therapy andnucleoside-based inhibitors of HBV DNA polymerase, namely entecavir andtenofovir. The current standard of care is dedicated to reducing thelevel of viremia and allowance of liver dysfunction, but is associatedwith negative side-effects and increase persistence of drug-resistantHBV mutants. A significant shortcoming of current therapies is that theyare unable to eliminate hepatic reservoirs of cccDNA, preventtranscription of HBsAg from cccDNA, or limit the secretion of HBsAg intoserum that will ultimately stifle the immune response. Althoughcompounds have been reported to reduce serum HBsAg levels, they have notbeen approved as HBV therapies. (Refer to WO2015/113990, WO2015/173164,WO2016/023877, WO2016/071215, WO2016/128335, WO 2017/140821,WO2019097479, WO2019166951, WO2019123285, WO2018198079, WO2018073753,WO2018047109, WO2019110352, WO2019129681, WO2018087345, WO2018083136,WO2018083106, WO2018083081, WO2017216391, WO2018001952, WO2018001944,WO2016107832, WO2016177655, WO2017017042, WO2017017043. WO2017013046,WO2018219356, WO2018130152, WO2018154466, WO2019069293, WO2017061466,WO2018181883, WO2018161960, WO2017205115, WO2018144605, WO2018085619,WO2018019297, and WO2018022282).

More effective therapies for chronic HBV infections are needed due tothis high unmet clinical need. This invention describes the methods toprepare and methods for use of compounds that are believed to suppressthe secretion of subviral particles containing HBsAg. Compounds of thistype might be used to treat HBV infections and decrease occurrence ofliver disease complications such as cirrhosis or HCC.

There is a need in the art for novel therapeutic agents that treat,ameliorate or prevent HBV infection. Administration of these therapeuticagents to an HBV infected patient, either as monotherapy or incombination with other HBV treatments or ancillary treatments, will leadto significantly improved prognosis, diminished progression of thedisease, and enhanced seroconversion rates.

SUMMARY OF THE INVENTION

The present invention relates to novel antiviral compounds,pharmaceutical compositions comprising such compounds, as well asmethods to treat or prevent viral (particularly HBV) infection in asubject in need of such therapy with said compounds. Compounds of thepresent invention inhibit the protein(s) encoded by hepatitis B virus(HBV) or interfere with the life cycle of HBV and are also useful asantiviral agents. In addition, the present invention provides processesfor the preparation of said compounds.

The present invention provides compounds represented by Formula (I),

and pharmaceutically acceptable salts, N-oxides, esters and prodrugsthereof, wherein:

Q₁, Q₂, Q₃, and Q₄ are each independently selected from hydrogen, halo,—NR₁₁R₁₂, optionally substituted —C₁-C₆ alkyl, optionally substituted—C₂-C₆ alkenyl, optionally substituted —C₁-C₆ alkoxy, optionallysubstituted —C₃-C₈ cycloalkyl; optionally substituted —C₃-C₈cycloalkenyl; optionally substituted 3- to 8-membered heterocycloalkyl;optionally substituted aryl; and optionally substituted heteroaryl;

Y₁ is hydrogen, halo, optionally substituted C₁-C₆ alkyl, optionallysubstituted —C₃-C₆ cycloalkyl, optionally substituted 3- to 6-memberedheterocycloalkyl, —CN, —C(O)R₁₁; —C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂);—C(O)N(R₁₁)S(O)₂(R₁₂); —OR₁₁, —NR₁₁R₁₂; —SR₁₁; —S(O)₂R₁₁; or—S(O)₂N(R₁₁)(R₁₂);

Y₂ is hydrogen, halo, optionally substituted C₁-C₆ alkyl, optionallysubstituted —C₃-C₆ cycloalkyl, optionally substituted 3- to 6-memberedheterocycloalkyl, —CN, —C(O)R₁₁; —C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂);—C(O)N(R₁₁)S(O)₂(R₁₂); —OR₁₁, —NR₁₁R₁₂; —SR₁₁; —S(O)₂R₁₁;—S(O)₂N(R₁₁)(R₁₂), or —O—Si(R₁₁)₃;

Y₃ is —C(O)R₁₁, —COOR₁₁, —C(O)NHSO₂R₁₁, —C(O)NHSO₂NR₁₁R₁₂, or1,2,4-oxadiazol-3-yl-5(4H)-one, or Y₃ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₅-C₆cycloalkyl, or optionally substituted 5- to 6-membered heterocycloalkyl;

Y₄ is hydrogen, halo, optionally substituted C₁-C₆ alkyl, optionallysubstituted —C₃-C₆ cycloalkyl, optionally substituted 3- to 6-memberedheterocycloalkyl, —CN, —C(O)R₁₁; —C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂);—C(O)N(R₁₁)S(O)₂(R₁₂); —OR₁₁, —NR₁₁R₁₂; —SR₁₁; —S(O)₂R₁₁; or—S(O)₂N(R₁₁)(R₁₂);

Alternatively, Y₂ and Y₃ are taken together to form an optionallysubstituted 5-12 membered heterocyclic ring or carbocyclic ringcontaining 1, 2, or 3 double bonds;

Z₁ is N or CR₁, Z₂ is N or CR₂, and Z₃ is N or CR₃; in certainembodiments, at least one of Z₁, Z₂ and Z₃ is N;

R₁, R₂ and R₃ are each independently selected from:

1) hydrogen;

2) halogen;

3) —NO₂;

4) Cyano;

5) Optionally substituted —C₁-C₈ alkyl;

6) Optionally substituted —C₂-C₈ alkenyl;

7) Optionally substituted —C₂-C₈ alkynyl;

8) Optionally substituted —C₃-C₈ cycloalkyl;

9) Optionally substituted 3- to 12-membered heterocycloalkyl;

10) Optionally substituted aryl;

11) Optionally substituted arylalkyl;

12) Optionally substituted heteroaryl;

13) Optionally substituted heteroarylalkyl;

14) —SR₁₁;

15) —S(O)₂R₁₁;

16) —S(O)₂N(R₁₁)(R₁₂);

17) —C(O)R₁₁;

18) —C(O)OR₁₁;

19) —C(O)N(R₁₁)(R₁₂);

20) —C(O)N(R₁₁)S(O)₂(R₁₂);

21) —N(R₁₁)(R₁₂);

22) —N(R₁₃)C(O)N(R₁₁)(R₁₂);

23) —N(R₁₁)C(O)(R₁₂);

24) —N(R₁₁)C(O)₂(R₁₂);

25) —N(R₁₃)S(O)₂N(R₁₁)(R₁₂);

26) —N(R₁₁)S(O)₂(R₁₂);

27) —OR₁₁;

28) —OC(O)R₁₁;

29) —OC(O)OR₁₁; and

30) —OC(O)N(R₁₁)(R₁₂);

wherein R₁₁, R₁₂, and R₁₃, are each independently selected fromhydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted—C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionallysubstituted 3- to 8-membered heterocycloalkyl, optionally substitutedaryl, and optionally substituted heteroaryl. Alternatively, R₁₁ and R₁₂are taken together with the nitrogen atom to which they attached to forman optionally substituted 3-8 membered heterocyclic containing 0, 1, 2,or 3 double bonds.

Preferably, R₂ is optionally substituted aryl, optionally substitutedheteroaryl or optionally substituted bicyclic heterocycloalkyl, morepreferably optionally substituted aryl or optionally substitutedheteroaryl.

Preferably R₁ is optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted —C₃-C₈ cycloalkyl or optionallysubstituted 3- to 12-membered heterocycloalkyl; and R₂ is optionallysubstituted aryl, optionally substituted heteroaryl, optionallysubstituted —C₃-C₈ cycloalkyl or optionally substituted 3- to12-membered heterocycloalky.

In certain embodiments, Z₃ is N, Z₁ is CR₁ and Z₂ is CR₂, and R₂ is nothydrogen; halogen; cyano; optionally substituted —C₁-C₆ alkyl;optionally substituted —C₃-C₇ cycloalkyl; optionally substituted 3- to7-membered heterocycloalkyl; —NH₂; —NHC₁-C₆ alkyl; —OH; or —OC₁-C₆alkyl. In this embodiment, R₂ is preferably optionally substituted aryl,optionally substituted heteroaryl, optionally substituted arylalkyl, oroptionally substituted heteroarylalkyl, and R₁ is preferably optionallysubstituted aryl, optionally substituted arylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl, hydrogenor halogen.

Each preferred group stated above can be taken in combination with one,any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment of the present invention is a compound of Formula (I)as described above, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Y₁is hydrogen, F, Cl, —NH₂, —CH₃, —CHF₂, —CF₃, —OCH₃, or optionallysubstituted cyclopropyl.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Y₂is F, Cl, —NH₂, —CH₃, —CHF₂, —CF₃, or —OCH₃.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Y₂is —OH.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Y₃is —COOH, —C(O)NHSO₂NR₁₁R₁₂, wherein R₁₁ and R₁₂ are as previouslydefined or Y₃ is triazolyl.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Y₄is hydrogen, —OH, or —CH₃.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein atleast one of R₁, R₂ and R₃, preferably R₂, is optionally substitutedaryl; optionally substituted arylalkyl; optionally substitutedheteroaryl; or optionally substituted heteroarylalkyl.

In certain embodiments, Z₃ is N, Z₂ is CR₂, and Z₁ is CR₁, wherein R₁and R₂ are as defined above. In certain embodiments, R₁ is hydrogen orhalogen, preferably hydrogen. In certain embodiments, R₂ is optionallysubstituted aryl; optionally substituted arylalkyl; optionallysubstituted heteroaryl; or optionally substituted heteroarylalkyl.

In certain embodiments, Z₃ is N, Z₁ is CR₁, and Z₂ is CR₂, wherein R₁ isoptionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, or optionallysubstituted 3- to 12-membered heterocycloalkyl; and R₂ is as previouslydefined. Preferably, R₁ is optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted —C₃-C₈ cycloalkyl, oroptionally substituted 3- to 8-membered heterocycloalkyl; and R₂ is aspreviously defined. More preferably, R₁ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl;and R₂ is hydrogen.

In certain embodiments, Z₃ is N, Z₁ is CR₁, and Z₂ is CR₂, wherein R₂ isoptionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, or optionallysubstituted 3- to 12-membered heterocycloalkyl; and R₁ is as previouslydefined. Preferably, R₂ is optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted —C₃-C₈ cycloalkyl, oroptionally substituted 3- to 8-membered heterocycloalkyl; and R₁ is aspreviously defined. More preferably, R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl;and R₁ is hydrogen.

In certain embodiments, Z₃ is N, Z₁ is CR₁, and Z₂ is CR₂, wherein R₁ isoptionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, or optionallysubstituted 3- to 12-membered heterocycloalkyl; and R₂ is optionallysubstituted aryl, optionally substituted arylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl,optionally substituted —C₃-C₈ cycloalkyl, or optionally substituted 3-to 12-membered heterocycloalkyl. Preferably, R₁ is optionallysubstituted aryl, optionally substituted heteroaryl, optionallysubstituted —C₃-C₈ cycloalkyl, or optionally substituted 3- to8-membered heterocycloalkyl; and R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl.More preferably, R₁ is optionally substituted aryl or optionallysubstituted heteroaryl; and R₂ is optionally substituted aryl oroptionally substituted heteroaryl.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein atleast one of R₁, R₂ and R₃, preferably at least one of R₁ and R₂, isderived from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to fourgroups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substitutedC₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein atleast one of R₁, R₂ and R₃, preferably at least one of R₁ and R₂, isselected from one of the following:

wherein each R₂₁ is independently selected from —CH₃, -isopropyl,-t-butyl, or one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to fourgroups selected from halo, CN, —OR₁₁, —NR₁₁ R₁₂, optionally substitutedC₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Q₁is hydrogen, Cl or F; Q₂ is hydrogen, Cl or F; Q₃ is optionallysubstituted C₁-C₆ alkyl, hydrogen, C₁ or F and Q₄ is optionallysubstituted C₁-C₆ alkyl, hydrogen, Cl or F.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Q₁is hydrogen; Q₂ is hydrogen; Q₃ is optionally substituted C₁-C₆ alkyl orhydrogen, and Q₄ is optionally substituted C₁-C₆ alkyl or hydrogen.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein Q₄is -t-butyl or isopropyl.

In certain embodiments, the compound of Formula (I) is represented byone of Formulae (II-1)˜(II-8), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, R₁, R₂, R₃, Q₁, Q₂, Q₃, and Q₄ are as previouslydefined.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (III-1)˜Formulae (III-8), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, Y₂, Y₄, R₁, R₂, R₃, R₁₁, Q₃ and Q₄ are as previouslydefined.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (III-1a)˜Formulae (III-8a), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, Y₂, Y₄, R₁, R₂, R₃, R₁₁, and Q₄ are as previously defined.

In another embodiment, the compound of Formula (I) is represented byFormula (IV-1) or Formula (IV-2), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, and Z₃ are as previously defined.

In another embodiment, the compound of Formula (I) is represented byFormula (IV-1a) or Formula (IV-2a), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, and Z₃ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (V-1)˜Formulae (V-8), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, R₁, and R₂ are as previously defined. Preferably, R₁ isoptionally substituted aryl, optionally substituted heteroaryl,optionally substituted —C₃-C₈ cycloalkyl, or optionally substituted 3-to 8-membered heterocycloalkyl; and R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (VI-1)˜Formulae (VI-8), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, Y₂, R₁, and R₂ are as previously defined. Preferably, R₁ isoptionally substituted aryl, optionally substituted heteroaryl,optionally substituted —C₃-C₈ cycloalkyl, or optionally substituted 3-to 8-membered heterocycloalkyl; and R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (VII-1a)˜Formulae (VII-8a), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, R₁, and R₂ are as previously defined. Preferably, R₁ isoptionally substituted aryl, optionally substituted heteroaryl,optionally substituted —C₃-C₈ cycloalkyl, or optionally substituted 3-to 8-membered heterocycloalkyl; and R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (VII-1)˜Formulae (VII-8), or a pharmaceutically acceptablesalt thereof:

wherein Y₁, R₁, and R₂ are as previously defined. Preferably, R₁ isoptionally substituted aryl, optionally substituted heteroaryl,optionally substituted —C₃-C₈ cycloalkyl, or optionally substituted 3-to 8-membered heterocycloalkyl; and R₂ is optionally substituted aryl,optionally substituted heteroaryl, optionally substituted —C₃-C₈cycloalkyl, or optionally substituted 3- to 8-membered heterocycloalkyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (VIII-1)˜(VIII-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, R₁, R₂, and R₃ are as previously defined. Preferably R₁,R₂, and R₃ are each independently selected from hydrogen, halogen,optionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, andoptionally substituted 3- to 8-membered heterocycloalkyl. Morepreferably, Y₁ is H, F or cycloproply; Y₂ is —OH; and R₁, R₂, and R₃ areeach independently selected from optionally substituted aryl andoptionally substituted heteroaryl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (IX-1)˜(IX-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, R₁, R₂, and R₃ are as previously defined. Preferably R₁,R₂, and R₃ are each independently selected from hydrogen, halogen,optionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, andoptionally substituted 3- to 8-membered heterocycloalkyl. Morepreferably, Y₁ is H, F, or cycloproply; Y₂ is —OH; and R₁, R₂, and R₃are each independently selected from optionally substituted aryl andoptionally substituted heteroaryl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (X-1)˜(X-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, R₁, R₂, and R₃ are as previously defined. Preferably R₁, R₂,and R₃ are each independently selected from hydrogen, halogen,optionally substituted aryl, optionally substituted arylalkyl,optionally substituted heteroaryl, optionally substitutedheteroarylalkyl, optionally substituted —C₃-C₈ cycloalkyl, andoptionally substituted 3- to 8-membered heterocycloalkyl. Morepreferably, Y₁ is H, F, or cycloproply; and R₁, R₂, and R₃ are eachindependently selected from optionally substituted aryl and optionallysubstituted heteroaryl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XI-1)˜(XI-8), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, R₁, R₂ and R₃ are as previously defined and R₁,R₂ and R₃ are not hydrogen. Preferably, Y₁ is H, F or cycloproply; Y₂ is—OH; Y₃ is —COOR₁₁; Y₄ is hydrogen; and R₁, R₂, and R₃ are eachindependently selected from optionally substituted aryl and optionallysubstituted heteroaryl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XII-1)˜(XII-8), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, R₁, R₂ and R₃ are as previously defined and R₁,R₂ and R₃ are not hydrogen. Preferably, Y₁ is H, F or cycloproply; Y₂ is—OH; Y₃ is —COOR₁₁; Y₄ is hydrogen; and R₁, R₂, and R₃ are eachindependently selected from optionally substituted aryl and optionallysubstituted heteroaryl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XIII-1)˜(XIII-6), or a pharmaceutically acceptable saltthereof:

wherein R₂₂ and R₂₃ are each independently selected from halo, CN,—OR₁₁, —N₁₁R₁₂, and optionally substituted C₁-C₆ alkyl; m is 0, 1, 2, 3or 4; n is 0, 1, 2, 3, or 4; Y₁, Y₂, Y₃, and Y₄ are as previouslydefined. Preferably, Y₁ is H, F or cycloproply; Y₂ is —OH; Y₃ is—COOR₁₁; and Y₄ is hydrogen.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XIV-1)˜(XIV-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, R₂₂, R₂₃, m and n are as previously defined. Preferably,Y₁ is H, F or cycloproply; and Y₂ is —OH.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XV-1)˜(XV-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, R₂₂, R₂₃, m and n are as previously defined.Preferably, Y₁ is H, F or cycloproply; Y₂ is —OH; Y₃ is —COOR₁₁; and Y₄is hydrogen.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XVI-1)˜(XVI-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, R₂₂, R₂₃, m and n are as previously defined. Preferably,Y₁ is H, F, or cycloproply; and Y₂ is —OH.

In another embodiment, the compound of the invention is represented byFormula (II-1)˜Formula (II-8), or Formula (III-1)˜Formula (III-8), orFormula (XI-1)˜(XI-8), or Formula (XII-1)˜Formula (XII-8), or apharmaceutically acceptable salt thereof, wherein at least one of R₁,R₂, and R₃ is independently selected from the groups in Table 1. Inanother embodiment, the compound of the invention is represented byFormula (V-1)˜Formula (V-8), or Formula (VI-1)˜Formula (VI-8), orFormula (VII-1)˜(VIII-8), or Formula (VII-1a)˜(VIII-8a), or Formula(VIII-1)˜Formula (VIII-6), or Formula (IX-1)˜Formula (IX-6), or Formula(X-1)˜Formula (X-6), or a pharmaceutically acceptable salt thereof,wherein at least one of R₁ and R₂ is independently selected from thegroups in Table 1.

TABLE 1 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

In another embodiment, the compound of Formula (I) is represented byFormula (XVII), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Q₁, and Q₂ are as previouslydefined.

In another embodiment, the compound of Formula (I) is represented byFormula (XVIIa) or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Q₁, and Q₂ are as previouslydefined. Preferably, Q₃ is hydrogen or optionally substituted C₁-C₆alkyl; and Q₄ is hydrogen, optionally substituted C₁-C₆ alkyl,optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈cycloalkyl; optionally substituted —C₃-C₅ cycloalkenyl; optionallysubstituted 3- to 8-membered heterocycloalkyl. More preferably, Q₃ isoptionally substituted C₁-C₆ alkyl or hydrogen; Q₄ is optionallysubstituted C₁-C₆ alkyl or hydrogen.

In another embodiment, the compound of Formula (I) is represented byFormula (XVIIa) or a pharmaceutically acceptable salt thereof, whereinY₁, Y₂, Y₃, Y₄, Z₁, Z₂, and Z₃ are as previously defined, Q₃ isoptionally substituted methyl and Q₄ is optionally substituted methyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XVIII-1)˜(XVIII-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Y₃, Y₄, Q₃, Q₄, R₂₂, R₂₃, m and n are as previouslydefined. Preferably, Q₃ is hydrogen or optionally substituted C₁-C₆alkyl; and Q₄ is hydrogen, optionally substituted C₁-C₆ alkyl,optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈cycloalkyl; optionally substituted —C₃-C₈ cycloalkenyl, or optionallysubstituted 3- to 8-membered heterocycloalkyl.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XIX-1)˜(XIX-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Y₂, Q₃, Q₄, R₂₂, R₂₃, m and n are as previously defined.

In another embodiment, the compound of Formula (I) is represented by oneof Formulae (XX-1)˜(XX-6), or a pharmaceutically acceptable saltthereof:

wherein Y₁, Q₃, Q₄, R₂₂, R₂₃, m and n are as previously defined.

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principles ofchemical bonding. In some instances, it may be necessary to remove ahydrogen atom in order to accommodate a substituent at any givenlocation.

It will be yet appreciated that the compounds of the present inventionmay contain one or more asymmetric carbon atoms and may exist inracemic, diastereoisomeric, and optically active forms. It will still beappreciated that certain compounds of the present invention may exist indifferent tautomeric forms. All tautomers are contemplated to be withinthe scope of the present invention.

In one embodiment, the compounds described herein are suitable formonotherapy and are effective against natural or native HBV strains andagainst HBV strains resistant to currently known drugs. In anotherembodiment, the compounds described herein are suitable for use incombination therapy.

In another embodiment, the compounds of the invention can be used inmethods of modulating (e.g., inhibit, disrupt or accelerate) theactivity of HBV cccDNA. In yet another embodiment, the compounds of theinvention can be used in methods of diminishing or preventing theformation of HBV cccDNA. In another embodiment, the additionaltherapeutic agent is selected from core inhibitor, which includes GLS4,GLS4JHS, JNJ-379, ABI-H0731, ABI-H2158, AB-423, AB-506, WX-066, andQL-0A6A; immune modulator or immune stimulator therapies, which includesT-cell response activator AIC649 and biological agents belonging to theinterferon class, such as interferon alpha 2a or 2b or modifiedinterferons such as pegylated interferon, alpha 2a, alpha 2b, lamda; orSTING (stimulator of interferon genes) modulator; or TLR modulators suchas TLR-7 agonists, TLR-8 agonists or TLR-9 agonists; or therapeuticvaccines to stimulate an HBV-specific immune response such as virus-likeparticles composed of HBcAg and HBsAg, immune complexes of HBsAg andHBsAb, or recombinant proteins comprising HBx, HBsAg and HBcAg in thecontext of a yeast vector; or immunity activator such as SB-9200 ofcertain cellular viral RNA sensors such as RIG-I, NOD2, and MDA5protein, or RNA interference (RNAi) or small interfering RNA (siRNA)such as ARC-520, ARC-521, ARB-1467, and ALN-HBV RNAi, or antiviralagents that block viral entry or maturation or target the HBV polymerasesuch as nucleoside or nucleotide or non-nucleos(t)ide polymeraseinhibitors, and agents of distinct or unknown mechanism including agentsthat disrupt the function of other essential viral protein(s) or hostproteins required for HBV replication or persistence such as REP 2139,RG7834, and AB-452. In an embodiment of the combination therapy, thereverse transcriptase inhibitor is at least one of Zidovudine,Didanosine, Zalcitabine, ddA, Stavudine, Lamivudine, Aba-cavir,Emtricitabine, Entecavir, Apricitabine, Atevirapine, ribavirin,acyclovir, famciclovir, valacyclovir, ganciclovir, valganciclovir,Tenofovir, Adefovir, PMPA, cidofovir, Efavirenz, Nevirapine,Delavirdine, or Etravirine.

In another embodiment of the combination therapy, the TLR-7 agonist isselected from the group consisting of SM360320(12-benzyl-8-hydroxy-2-(2-methoxy-ethoxy)ad-enine), AZD 8848 (methyl[3-({[3-(6-amino-2-butoxy-8-oxo-7,8-dihydro-9H-purin-12-yl)propyl][3-(4-morpholinyl)propyl] amino Imethyl)phenyl] acetate), GS-9620(4-Amino-2-butoxy-8-[3-(2-pyrrolidinylmethyl)benzyl]-7,8-dihydro-6(5H)-pteridinone),AL-034 (TQ-A3334), and RO6864018.

In another embodiment of the combination therapy, the TLR-8 agonist isGS-9688.

In an embodiment of these combination therapies, the compound and theadditional therapeutic agent are co-formulated. In another embodiment,the compound and the additional therapeutic agent are co-administered.

In another embodiment of the combination therapy, administering thecompound of the invention allows for administering of the additionaltherapeutic agent at a lower dose or frequency as compared to theadministering of the at least one additional therapeutic agent alonethat is required to achieve similar results in prophylactically treatingan HBV infection in an individual in need thereof.

In another embodiment of the combination therapy, before administeringthe therapeutically effective amount of the compound of the invention,the individual is known to be refractory to a compound selected from thegroup consisting of a HBV polymerase inhibitor, interferon, viral entryinhibitor, viral maturation inhibitor, distinct capsid assemblymodulator, antiviral compounds of distinct or unknown mechanism, andcombination thereof.

In still another embodiment of the method, administering the compound ofthe invention reduces viral load in the individual to a greater extentcompared to the administering of a compound selected from the groupconsisting of a HBV polymerase inhibitor, interferon, viral entryinhibitor, viral maturation inhibitor, distinct capsid assemblymodulator, antiviral compounds of distinct or unknown mechanism, andcombination thereof.

In another embodiment, administering of the compound of the inventioncauses a lower incidence of viral mutation and/or viral resistance thanthe administering of a compound selected from the group consisting of aHBV polymerase inhibitor, interferon, viral entry inhibitor, viralmaturation inhibitor, distinct capsid assembly modulator, antiviralcompounds of distinct or unknown mechanism, and combination thereof.

It should be understood that the compounds encompassed by the presentinvention are those that are suitably stable for use as pharmaceuticalagent.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono- or polycycliccarbocyclic ring system comprising at least one aromatic ring,including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl,indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system thatcomprises at least one aromatic ring. Polycyclic aryls can comprisefused rings, covalently attached rings or a combination thereof.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclicaromatic radical having one or more ring atom selected from S, O and N;and the remaining ring atoms are carbon, wherein any N or S containedwithin the ring may be optionally oxidized. Heteroaryl includes, but isnot limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzoxazolyl, quinoxalinyl. A polycyclic heteroaryl can comprise fusedrings, covalently attached rings or a combination thereof.

In accordance with the invention, aromatic groups can be substituted orunsubstituted.

The term “alkyl” as used herein, refers to saturated, straight- orbranched-chain hydrocarbon radicals. “C₁-C₄ alkyl,” “C₁-C₆ alkyl,”“C₁-C₈ alkyl,” “C₁-C₁₂ alkyl,” “C₂-C₄ alkyl,” or “C₃-C₆ alkyl,” refer toalkyl groups containing from one to four, one to six, one to eight, oneto twelve, 2 to 4 and 3 to 6 carbon atoms respectively. Examples ofC₁-C₈ alkyl radicals include, but are not limited to, methyl, ethyl,propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl andoctyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon double bond bythe removal of a single hydrogen atom. “C₂-C₅ alkenyl,” “C₂-C₁₂alkenyl,” “C₂-C₄ alkenyl,” “C₃-C₄ alkenyl,” or “C₃-C₆ alkenyl,” refer toalkenyl groups containing from two to eight, two to twelve, two to four,three to four or three to six carbon atoms respectively. Alkenyl groupsinclude, but are not limited to, for example, ethenyl, propenyl,butenyl, 2-methyl-2-buten-2-yl, heptenyl, octenyl, and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon double bond bythe removal of a single hydrogen atom. “C₂-C₈ alkynyl,” “C₂-C₁₂alkynyl,” “C₂-C₄ alkynyl,” “C₃-C₄ alkynyl,” or “C₃-C₆ alkynyl,” refer toalkynyl groups containing from two to eight, two to twelve, two to four,three to four or three to six carbon atoms respectively. Representativealkynyl groups include, but are not limited to, for example, ethynyl,2-propynyl, 2-butynyl, heptynyl, octynyl, and the like.

The term “cycloalkyl”, as used herein, refers to a monocyclic orpolycyclic saturated carbocyclic ring or a bi- or tri-cyclic groupfused, bridged or spiro system, and the carbon atoms may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic doublebond. Preferred cycloalkyl groups include C₃-C₁₂ cycloalkyl, C₃-C₆cycloalkyl, C₃-C₈ cycloalkyl and C₄-C₇ cycloalkyl. Examples of C₃-C₁₂cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl,4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl,spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, andthe like.

The term “cycloalkenyl”, as used herein, refers to monocyclic orpolycyclic carbocyclic ring or a bi- or tri-cyclic group fused, bridgedor spiro system having at least one carbon-carbon double bond and thecarbon atoms may be optionally oxo-substituted or optionally substitutedwith exocyclic olefinic double bond. Preferred cycloalkenyl groupsinclude C₃-C₁₂ cycloalkenyl, C₃-C₈ cycloalkenyl or C₅-C₇ cycloalkenylgroups. Examples of C₃-C₁₂ cycloalkenyl include, but not limited to,cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl,cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl,spiro[2.5]oct-4-enyl, spiro[4.4]non-2-enyl,bicyclo[4.2.1]non-3-en-12-yl, and the like.

As used herein, the term “arylalkyl” means a functional group wherein analkylene chain is attached to an aryl group, e.g., —CH₂CH₂-phenyl. Theterm “substituted arylalkyl” means an arylalkyl functional group inwhich the aryl group is substituted. Similarly, the term“heteroarylalkyl” means a functional group wherein an alkylene chain isattached to a heteroaryl group. The term “substituted heteroarylalkyl”means a heteroarylalkyl functional group in which the heteroaryl groupis substituted.

As used herein, the term “alkoxy” employed alone or in combination withother terms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms connected to the rest of the moleculevia an oxygen atom, such as, for example, methoxy, ethoxy, 2-propoxy,2-propoxy (isopropoxy) and the higher homologs and isomers. Preferredalkoxy are (C₂-C₃) alkoxy.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclic and cycloalkenyl moiety described herein can also be analiphatic group or an alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of anycombination of carbon atoms, hydrogen atoms, halogen atoms, oxygen,nitrogen or other atoms, and optionally contains one or more units ofunsaturation, e.g., double and/or triple bonds. Examples of aliphaticgroups are functional groups, such as alkyl, alkenyl, alkynyl, O, OH,NH, NH₂, C(O), S(O)₂, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH,S(O)₂NH₂, NHC(O)NH₂, NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂,C(O)NHS(O)₂NH or C(O)NHS(O)₂NH₂, and the like, groups comprising one ormore functional groups, non-aromatic hydrocarbons (optionallysubstituted), and groups wherein one or more carbons of a non-aromatichydrocarbon (optionally substituted) is replaced by a functional group.Carbon atoms of an aliphatic group can be optionally oxo-substituted. Analiphatic group may be straight chained, branched, cyclic, or acombination thereof and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, as used herein, aliphaticgroups expressly include, for example, alkoxyalkyls, polyalkoxyalkyls,such as polyalkylene glycols, polyamines, and polyimines, for example.Aliphatic groups may be optionally substituted.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused, bridged or spiro system, where (i) each ringsystem contains at least one heteroatom independently selected fromoxygen, sulfur and nitrogen, (ii) each ring system can be saturated orunsaturated (iii) the nitrogen and sulfur heteroatoms may optionally beoxidized, (iv) the nitrogen heteroatom may optionally be quaternized,(v) any of the above rings may be fused to an aromatic ring, and (vi)the remaining ring atoms are carbon atoms which may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic doublebond. Representative heterocycloalkyl groups include, but are notlimited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl,imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl,isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl,quinoxalinyl, pyridazinonyl, 2-azabicyclo[2.2.1]-heptyl,8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl,2-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl, and tetrahydrofuryl.Such heterocyclic groups may be further substituted. Heteroaryl orheterocyclic groups can be C-attached or N-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphaticmoiety or the like, described herein can also be a divalent ormultivalent group when used as a linkage to connect two or more groupsor substituents, which can be at the same or different atom(s). One ofskill in the art can readily determine the valence of any such groupfrom the context in which it occurs.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms with substituentsincluding, but not limited to, —F, —Cl, —Br, —I, —OH, C₁-C₁₂-alkyl;C₂-C₁₂-alkenyl, C₂-C₁₂-alkynyl, —C₃-C₁₂-cycloalkyl, protected hydroxy,—NO₂, —N₃, —CN, —NH₂, protected amino, oxo, thioxo, —NH—C₁-C₁₂-alkyl,—NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl, —NH—C₃-C₁₂-cycloalkyl, —NH-aryl,—NH-heteroaryl, —NH— heterocycloalkyl, -dialkylamino, -diarylamino,-diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl,—O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O— heterocycloalkyl,—C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl,—C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)-heteroaryl,—C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl,—CONH—C₂-C₈-alkynyl, —CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl,—CONH-heteroaryl, —CONH-heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl,—OCO₂—C₂-C₈-alkenyl, —OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl,—OCO₂-aryl, —OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —CO₂—C₁-C₁₂ alkyl,—CO₂—C₂-C₅ alkenyl, —CO₂—C₂-C₈ alkynyl, CO₂—C₃-C₁₂-cycloalkyl, —CO₂—aryl, CO₂-heteroaryl, CO₂— heterocyloalkyl, —OCONH₂,—OCONH—C₁-C₁₂-alkyl, —OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl,—OCONH—C₃-C₁₂-cycloalkyl, —OCONH-aryl, —OCONH-heteroaryl,—OCONH-heterocyclo-alkyl, —NHC(O)H, —NHC(O)—C₁-C₁₂-alkyl,—NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl, —NHC(O)—C₃-C₁₂-cycloalkyl,—NHC(O)-aryl, —NHC(O)-heteroaryl, —NHC(O)-heterocyclo-alkyl,—NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl, —NHCO₂—C₂-C₈-alkynyl,—NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl, —NHCO₂-heteroaryl,—NHCO₂-heterocycloalkyl, —NHC(O)NH₂, —NHC(O)NH—C₁-C₁₂-alkyl,—NHC(O)NH—C₂-C₈-alkenyl, —NHC(O)NH—C₂-C₈-alkynyl,—NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl, —NHC(O)NH-heteroaryl,—NHC(O)NH-heterocycloalkyl, NHC(S)NH₂, —NHC(S)NH—C₁-C₁₂-alkyl,—NHC(S)NH—C₂-C₈-alkenyl, —NHC(S)NH—C₂-C₈-alkynyl,—NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl, —NHC(S)NH-heteroaryl,—NHC(S)NH— heterocycloalkyl, —NHC(NH)NH₂, —NHC(NH)NH—C₁-C₁₂-alkyl,—NHC(NH)NH—C₂-C₈-alkenyl, —NHC(NH)NH—C₂-C₈-alkynyl,—NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl,—NHC(NH)NH-heterocycloalkyl, —NHC(NH)—C₁-C₁₂-alkyl,—NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl, —S(O)—heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH— C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH— heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₂-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthio-methyl. In certain embodiments, the substituents areindependently selected from halo, preferably C₁ and F; C₁-C₄-alkyl,preferably methyl and ethyl; halo-C₁-C₄-alkyl, such as fluoromethyl,difluoromethyl, and trifluoromethyl; C₂-C₄-alkenyl; halo-C₂-C₄-alkenyl;C₃-C₆-cycloalkyl, such as cyclopropyl; C₁-C₄-alkoxy, such as methoxy andethoxy; halo-C₁-C₄-alkoxy, such as fluoromethoxy, difluoromethoxy, andtrifluoromethoxy, —CN; —OH; NH₂; C₁-C₄-alkylamino; di(C₁-C₄-alkyl)amino;and NO₂. It is understood that the aryls, heteroaryls, alkyls, and thelike can be further substituted. In some cases, each substituent in asubstituted moiety is additionally optionally substituted with one ormore groups, each group being independently selected from C₁-C₄-alkyl;—CF₃, —OCH₃, —OCF₃, —F, —C₁, —Br, —I, —OH, —NO₂, —CN, and —NH₂.Preferably, a substituted alkyl group is substituted with one or morehalogen atoms, more preferably one or more fluorine or chlorine atoms.

The term “halo” or halogen” alone or as part of another substituent, asused herein, refers to a fluorine, chlorine, bromine, or iodine atom.

The term “optionally substituted”, as used herein, means that thereferenced group may be substituted or unsubstituted. In one embodiment,the referenced group is optionally substituted with zero substituents,i.e., the referenced group is unsubstituted. In another embodiment, thereferenced group is optionally substituted with one or more additionalgroup(s) individually and independently selected from groups describedherein.

The term “hydrogen” includes hydrogen and deuterium. In addition, therecitation of an atom includes other isotopes of that atom so long asthe resulting compound is pharmaceutically acceptable.

The term “hydroxy activating group,” as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxyl,” as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl,chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl,methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl,benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl,benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl,trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy prodrug group,” as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992).

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, methoxycarbonyl, t-butoxycarbonyl, 12-fluorenyl-methoxycarbonyl,benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The term “protic solvent,” as used herein, refers to a solvent thattends to provide protons, such as an alcohol, for example, methanol,ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Suchsolvents are well known to those skilled in the art, and it will beobvious to those skilled in the art that individual solvents or mixturesthereof may be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable,” as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject,” as used herein, refers to an animal. Preferably, theanimal is a mammal. More preferably, the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

As used herein, the term “pharmaceutically acceptable salt,” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 2-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentane-propionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intra-arterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose, any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectable.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to Van Devanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of whichare incorporated herein by reference).

Combination and Alternation Therapy

It has been recognized that drug-resistant variants of HIV, HBV and HCVcan emerge after prolonged treatment with an antiviral agent. Drugresistance most typically occurs by mutation of a gene that encodes fora protein such as an enzyme used in viral replication, and mosttypically in the case of HIV, reverse transcriptase, protease, or DNApolymerase, and in the case of HBV, DNA polymerase, or in the case ofHCV, RNA polymerase, protease, or helicase. Recently, it has beendemonstrated that the efficacy of a drug against HIV infection can beprolonged, augmented, or restored by administering the compound incombination or alternation with a second, and perhaps third, antiviralcompound that induces a different mutation from that caused by theprinciple drug. The compounds can be used for combination are selectedfrom the group consisting of a HBV polymerase inhibitor, interferon, TLRmodulators such as TLR-7 agonists or TLR-9 agonists, therapeuticvaccines, immune activator of certain cellular viral RNA sensors, viralentry inhibitor, viral maturation inhibitor, distinct capsid assemblymodulator, antiviral compounds of distinct or unknown mechanism, andcombination thereof. Alternatively, the pharmacokinetics,biodistribution, or other parameter of the drug can be altered by suchcombination or alternation therapy. In general, combination therapy istypically preferred over alternation therapy because it induces multiplesimultaneous stresses on the virus.

Preferred compounds for combination or alternation therapy for thetreatment of HBV include 3TC, FTC, L-FMAU, interferon, adefovirdipivoxil, entecavir, telbivudine (L-dT), valtorcitabine (3′-valinylL-dC), β-D-dioxolanyl-guanine (DXG), β-D-dioxolanyl-2,6-diaminopurine(DAPD), and β-D-dioxolanyl-6-chloropurine (ACP), famciclovir,penciclovir, lobucavir, ganciclovir, and ribavirin.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

Antiviral Activity

An inhibitory amount or dose of the compounds of the present inventionmay range from about 0.01 mg/Kg to about 500 mg/Kg, alternatively fromabout 1 to about 50 mg/Kg. Inhibitory amounts or doses will also varydepending on route of administration, as well as the possibility ofco-usage with other agents.

According to the methods of treatment of the present invention, viralinfections, conditions are treated or prevented in a patient such as ahuman or another animal by administering to the patient atherapeutically effective amount of a compound of the invention, in suchamounts and for such time as is necessary to achieve the desired result.

By a “therapeutically effective amount” of a compound of the inventionis meant an amount of the compound which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of the compounddescribed above may range from about 0.1 mg/Kg to about 500 mg/Kg,preferably from about 1 to about 50 mg/Kg. Effective doses will alsovary depending on route of administration, as well as the possibility ofco-usage with other agents. It will be understood, however, that thetotal daily usage of the compounds and compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

The compounds of the present invention described herein can, forexample, be administered by injection, intravenously, intra-arterial,subdermally, intraperitoneally, intramuscularly, or subcutaneously; ororally, buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.1 toabout 500 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with pharmaceutically excipients or carriers toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations may contain from about 20% to about 80% activecompound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

When the compositions of this invention comprise a combination of acompound of the Formula described herein and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 1 to 100%, andmore preferably between about 5 to 95% of the dosage normallyadministered in a monotherapy regimen. The additional agents may beadministered separately, as part of a multiple dose regimen, from thecompounds of this invention. Alternatively, those agents may be part ofa single dosage form, mixed together with the compounds of thisinvention in a single composition.

The “additional therapeutic or prophylactic agents” include but are notlimited to, immune therapies (e.g. interferon), therapeutic vaccines,antifibrotic agents, anti-inflammatory agents such as corticosteroids orNSAIDs, bronchodilators such as beta-2 adrenergic agonists and xanthines(e.g. theophylline), mucolytic agents, anti-muscarinics,anti-leukotrienes, inhibitors of cell adhesion (e.g. ICAM antagonists),anti-oxidants (e.g. N-acetylcysteine), cytokine agonists, cytokineantagonists, lung surfactants and/or antimicrobial and anti-viral agents(e.g. ribavirin and amantidine). The compositions according to theinvention may also be used in combination with gene replacement therapy.

ABBREVIATIONS

Abbreviations which may be used in the descriptions of the scheme andthe examples that follow are: Ac for acetyl; AcOH for acetic acid; Boc₂Ofor di-tert-butyl-dicarbonate; Boc for t-butoxycarbonyl; Bz for benzoyl;Bn for benzyl; t-BuOK for potassium tert-butoxide; Brine for sodiumchloride solution in water; CDI for carbonyldiimidazole; DCM or CH₂Cl₂for dichloromethane; CH₃ for methyl; CH₃CN for acetonitrile; Cs₂CO₃ forcesium carbonate; CuCl for copper (I) chloride; CuI for copper (I)iodide; dba for dibenzylidene acetone; DBU for1,8-diazabicyclo[5.4.0]-undec-7-ene; DEAD for diethylazodicarboxylate;DIAD for diisopropyl azodicarboxylate; DIPEA or (i-Pr)₂EtN forN,N,-diisopropylethyl amine; DMP or Dess-Martin periodinane for1,1,2-tris(acetyloxy)-1,2-dihydro-1,2-benziodoxol-3-(1H)-one; DMAP for4-dimethylamino-pyridine; DME for 1,2-dimethoxyethane; DMF forN,N-dimethylformamide; DMSO for dimethyl sulfoxide; EtOAc for ethylacetate; EtOH for ethanol; Et₂O for diethyl ether; HATU forO-(7-azabenzotriazol-2-yl)-N,N,N′,N′,-tetramethyluroniumHexafluoro-phosphate; HCl for hydrogen chloride; K₂CO₃ for potassiumcarbonate; n-BuLi for n-butyl lithium; DDQ for2,3-dichloro-5,6-dicyano-1,4-benzoquinone; LDA for lithiumdiisopropylamide; LiTMP for lithium 2,2,6,6-tetramethyl-piperidinate;MeOH for methanol; Mg for magnesium; MOM for methoxymethyl; Ms for mesylor —SO₂—CH₃; NaHMDS for sodium bis(trimethylsilyl)amide; NaCl for sodiumchloride; NaH for sodium hydride; NaHCO₃ for sodium bicarbonate orsodium hydrogen carbonate; Na₂CO₃ sodium carbonate; NaOH for sodiumhydroxide; Na₂SO₄ for sodium sulfate; NaHSO₃ for sodium bisulfite orsodium hydrogen sulfite; Na₂S₂O₃ for sodium thiosulfate; NH₂NH₂ forhydrazine; NH₄Cl for ammonium chloride; Ni for nickel; OH for hydroxyl;OsO₄ for osmium tetroxide; OTf for triflate; PPA for polyphophoric acid;PTSA for p-toluenesulfonic acid; PPTS for pyridinium p-toluenesulfonate;TBAF for tetrabutylammonium fluoride; TEA or Et₃N for triethylamine; TESfor triethylsilyl; TESCl for triethylsilyl chloride; TESOTf fortriethylsilyl trifluoromethanesulfonate; TFA for trifluoroacetic acid;THE for tetrahydrofuran; TMEDA forN,N,N′,N′-tetramethylethylene-diamine; TPP or PPh₃ fortriphenyl-phosphine; Tos or Ts for tosyl or —SO₂—C₆H₄CH₃; Ts₂O fortolylsulfonic anhydride or tosyl-anhydride; TsOH for p-tolylsulfonicacid; Pd for palladium; Ph for phenyl; Pd₂(dba)₃ fortris(diben-zylideneacetone) dipalladium (0); Pd(PPh₃)₄ fortetrakis(triphenylphosphine)-palladium (0); PdCl₂(PPh₃)₂ fortrans-dichlorobis-(triphenylphosphine)palladium (II); Pt for platinum;Rh for rhodium; rt for room temperature; Ru for ruthenium; TBS fortert-butyl dimethylsilyl; TMS for trimethylsilyl; or TMSCl fortrimethylsilyl chloride.

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared. These schemes are of illustrative purpose, and are not meantto limit the scope of the invention. Equivalent, similar, or suitablesolvents, reagents or reaction conditions may be substituted for thoseparticular solvents, reagents, or reaction conditions described hereinwithout departing from the general scope of the method of synthesis.

Illustrated in Scheme 1, compounds such as 4 (Z₁, Z₂, Z₃, Q₁, Q₂, Q₃,and Q₄ as defined previously) can be prepared according to theillustrated synthetic methods herein, or by similar methods known tothose skilled in the art. Intermediate 1 can be reacted in acarbon-nitrogen bond forming reaction with ester 2 (M defined ashalogen, OMs, OAc, OTf, OTs, or ONs; Ru as defined previously),typically mediated by a base (denoted as [Base]) including, but notlimited to: K₂CO₃, Cs₂CO₃, KOAc, NaOtBu, NaOH, KOH, NaH, Et₃N, or DBU.Intermediate 3 can be reacted in a ring-closing step, typically mediatedby a base (denoted as [Base]) including, but not limited to: nBuLi orsBuLi, to produce ketone 4.

Illustrated in Scheme 2, compounds such as 5 (Z₁, Z₂, Z₃, Q₁, Q₂, Q₃,Q₄, Y₁, Y₂, Y₃, and Y₄ as defined previously) can be prepared accordingto the illustrated synthetic methods herein, or by similar methods knownto those skilled in the art. Ketone 1 can be reacted in a homologationreaction with orthoester 2 (R₁₁ as defined previously), typicallymediated by a base (denoted as [Base]) including, but not limited to:K₂CO₃, Cs₂CO₃, DIPEA, Et₃N, or DBU and an activator (denoted as[Activator]) including, but not limited to: BF₃·OEt₂, Ti(OiPr)₄, orB(C₆F₅)₃. Intermediate 3 can be reacted in an annulation reaction withalkene 4, which is first deprotonated using a base (denoted as [Base])including, but not limited to: LiHMDS, LDA, or LiTMP, then protectedusing an electrophilic reagent (denoted as [PG]) including, but notlimited to: TMSCl, TMSOTf, or TESOTf, then reacted with an activator(denoted as [Activator]) including, but not limited to: TiCl₄ orBF₃·OEt₂, to produce compound 5.

Illustrated in Scheme 3, compounds such as 3 (Z₂, Z₃, R₁, Q₁, Q₂, Q₃,Q₄, Y₁, Y₂, Y₃, and Y₄ as defined previously) can be prepared accordingto the illustrated synthetic methods herein, or by similar methods knownto those skilled in the art. Compound 1 (X defined as halogen, —B(OH)₂,—BF₃K, —B(pin), —OTf, or —OMs) can be reacted in a coupling reactionwith 2 (M defined as halogen, —B(OH)₂, —BF₃K, —B(pin), —OTf, or —OMs)that is mediated by a metal-containing reagent (denoted as [Metal])including, but not limited to reagents that contain: Pd, Cu, Zn, Fe, Ir,Ru, Rh, or Ni, and a base (denoted as [Base]) including, but not limitedto: KOAc, K₂CO₃, Cs₂CO₃, Et₃N, or K₃PO₄ to produce 3.

Illustrated in Scheme 4, compounds such as 3 (Z₁, Z₃, R₂, Q₁, Q₂, Q₃,Q₄, Y₁, Y₂, Y₃, and Y₄ as defined previously) can be prepared accordingto the illustrated synthetic methods herein, or by similar methods knownto those skilled in the art. Compound 1 (X defined as halogen, —B(OH)₂,—BF₃K, —B(pin), —OTf, or —OMs) can be reacted in a coupling reactionwith 2 (M defined as halogen, —B(OH)₂, —BF₃K, —B(pin), —OTf, or —OMs)that is mediated by a metal-containing reagent (denoted as [Metal])including, but not limited to reagents that contain: Pd, Cu, Zn, Fe, Ir,Ru, Rh, or Ni, and a base (denoted as [Base]) including, but not limitedto: KOAc, K₂CO₃, Cs₂CO₃, Et₃N, or K₃PO₄ to produce 3.

Illustrated in Scheme 5, compounds such as 2 (Z₁, Z₂, Z₃, Q₁, Q₂, Q₃,Q₄, Y₁, Y₂, and Y₄ as defined previously) can be prepared according tothe illustrated synthetic methods herein, or by similar methods known tothose skilled in the art. Intermediate 1 (R₁₁ as defined previously) canbe reacted under hydrolytic conditions mediated by NaOH, and followed bypurification using SFC, compounds such as 2 can be obtained with highoptical purity.

Illustrated in Scheme 6, compounds such as 7 (Q₁, Q₂, Q₃, Q₄, and R₁₁ asdefined previously; M defined as halogen, OMs, OAc, OTf, OTs, or ONs)can be prepared according to the illustrated synthetic methods herein,or by similar methods known to those skilled in the art. Carbamate 1(R₁₂ as defined previously) can be reacted with acid chloride 2 toproduce intermediate 3. This can undergo an alkylation reaction withester 4 (X defined as halogen, —OTf, —OTs, or —OMs) and a baseincluding, but not limited to: LDA, LiHMDS, or NaH, to produceintermediate 5. This can undergo a deprotection reaction (denoted as[Deprotection]) to produce acid 6. Following net reduction (denoted as[Reduction]) and functionalization (denoted a [Functionalization]),compound 7 is produced.

All references cited herein, whether in print, electronic, computerreadable storage media or other form, are expressly incorporated byreference in their entirety, including but not limited to, abstracts,articles, journals, publications, texts, treatises, internet web sites,databases, patents, and patent publications.

Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Starting materials were either available from a commercial vendor orproduced by methods well known to those skilled in the art.

General Conditions:

Mass spectra were run on LC-MS systems using electrospray ionization.These were Agilent 1290 Infinity II systems with an Agilent 6120Quadrupole detector. Spectra were obtained using a ZORBAX EclipseXDB-C18 column (4.6×30 mm, 1.8 micron). Spectra were obtained at 298Kusing a mobile phase of 0.1% formic acid in water (A) and 0.1% formicacid in acetonitrile (B). Spectra were obtained with the followingsolvent gradient: 5% (B) from 0-1.5 min, 5-95% (B) from 1.5-4.5 min, and95% (B) from 4.5-6 min. The solvent flowrate was 1.2 mL/min. Compoundswere detected at 210 nm and 254 nm wavelengths. [M+H]⁺ refers tomono-isotopic molecular weights.

NMR spectra were run on a Bruker 400 MHz spectrometer. Spectra weremeasured at 298K and referenced using the solvent peak. Chemical shiftsfor ¹H NMR are reported in parts per million (ppm).

Compounds were purified via reverse-phase high-performance liquidchromatography (RPHPLC) using a Gilson GX-281 automated liquid handlingsystem. Compounds were purified on a Phenomenex Kinetex EVO C18 column(250×21.2 mm, 5 micron), unless otherwise specified. Compounds werepurified at 298K using a mobile phase of water (A) and acetonitrile (B)using gradient elution between 0% and 100% (B), unless otherwisespecified. The solvent flowrate was 20 mL/min and compounds weredetected at 254 nm wavelength.

Alternatively, compounds were purified via normal-phase liquidchromatography (NPLC) using a Teledyne ISCO Combiflash purificationsystem. Compounds were purified on a REDISEP silica gel cartridge.Compounds were purified at 298K and detected at 254 nm wavelength.

Ex1: Synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

Step 1: Into a 10-L 4-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed THE (4500.00 mL), LiCl(171.00 g, 4033.590 mmol, 1.59 equiv), TEA (641.00 g, 6334.61 mmol, 2.49equiv). This was followed by the addition of 2,2-dimethylpropanoylchloride (357.00 g, 2960.69 mmol, 1.17 equiv) dropwise with stirring at0° C. in 10 min. To this was added a solution of 3,3-dimethylbutanoicacid (354.00 g, 3047.521 mmol, 1.20 equiv) in THE (1500 mL) dropwisewith stirring at 0° C. in 30 min. To the mixture was added(4R)-4-benzyl-1,3-oxazolidin-2-one (450.00 g, 2539.46 mmol, 1.00 equiv),DMAP (15.51 g, 126.97 mmol, 0.05 equiv) at 0° C. The resulting solutionwas stirred for 1 overnight at room temperature. The reaction was thenquenched by the addition of 5 L of water. The resulting solution wasextracted with 2×3 L of ethyl acetate. The combined organic phase waswashed with 1×3 L of saturated aqueous NaHCO₃. The resulting mixture waswashed with 1×3 L of brine. The mixture was dried over anhydrous sodiumsulfate and concentrated under vacuum. The crude product was purified byre-crystallization from ether. This resulted in 590 g (84.38%) of(4R)-4-benzyl-3-(3,3-dimethylbutanoyl)-1,3-oxazolidin-2-one as a whitesolid.

Step 2: Into a 20-L 4-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed THE (5900.00 mL),diisopropylamine (345.80 g, 3423.76 mmol, 1.60 equiv). This was followedby the addition of n-BuLi (1284.00 mL, 3214.11 mmol, 2.50 equiv, 2.5 M)drop wise with stirring at −20° C. in 30 min. The resulting solution wasstirred for 30 min at −20° C. To this was added a solution of(4R)-4-benzyl-3-(3,3-dimethylbutanoyl)-1,3-oxazolidin-2-one (590.00 g,2142.74 mmol, 1.00 equiv) in THE (2000 mL) dropwise with stirring at−78° C. in 30 min. The resulting solution was stirred at −40° C. for 2.5h. To the mixture was added tert-butyl 2-bromoacetate (832.00 g, 4265.44mmol, 1.99 equiv) drop wise with stirring at −78° C. in 20 min. Theresulting mixture was warmed to room temperature slowly. The resultingsolution was stirred for 30 min at room temperature. The reaction wasthen quenched by the addition of 200 g of AcOH at 0° C. The resultingsolution was diluted with 5 L of H₂O. The resulting solution wasextracted with 2×5 L of ethyl acetate. The combined organic phase waswashed with 2×5 L of brine. The mixture was dried over anhydrous sodiumsulfate and concentrated under vacuum. The residue was applied onto asilica gel column with ethyl acetate/petroleum ether (1:10). Thisresulted in 335 g (40.14%) of tert-butyl(3R)-3-[(4R)-4-benzyl-2-oxo-1,3-oxazolidine-3-carbonyl]-4,4-dimethylpentanoateas a white solid.

Step 3: Into a 20-L 4-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed THE (7290.00 mL), H₂O(2430.00 mL), H₂O₂ (314.60 g, 2772.84 mmol, 8.00 equiv, 30%), LiOH(33.20 g, 1386.42 mmol, 4.00 equiv), tert-butyl(3R)-3-[(4R)-4-benzyl-2-oxo-1,3-oxazolidine-3-carbonyl]-4,4-dimethylpentanoate(135.00 g, 346.605 mmol, 1.00 equiv). The resulting solution was stirredfor 3 days at 30° C. The reaction was then quenched by the addition of2776 mL of Na₂SO₃ (1N). The resulting solution was extracted with 2×5 Lof ethyl acetate. The combined organic phase was washed with 2×5 L ofbrine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:5). The crude product (32g) was purified by silica gel chromatography to produce 22.9 g (28.69%)of (2R)-2-[2-(tert-butoxy)-2-oxoethyl]-3,3-dimethylbutanoic acid as alight yellow semi-solid. ESI MS m/z=229.1 [M−H]⁻. H-NMR (400 MHz, CDCl₃,ppm): δ 2.747-2.592 (2H, m), 2.463-2.406 (1H, m), 1.449 (9H, s), 1.020(9H, s).

Step 4: Into a 2-L 4-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-3,3-dimethylbutanoic acid (52.00 g,225.78 mmol, 1.00 equiv). This was followed by the addition of THF(500.00 mL). To this was added BH₃-Me₂S (45.16 mL, 2.00 equiv, 10 M)drop wise with stirring at 0-10° C. in 20 min. The resulting solutionwas stirred for overnight at room temperature. The reaction was thenquenched by the addition of 500 mL of water at −10° C. The resultingsolution was extracted with 2×200 mL of ethyl acetate and the organiclayers combined. The solids were filtered out. The resulting mixture waswashed with 300 mL of saturated aqueous NaHCO₃. The resulting mixturewas washed with 300 mL of H₂O. The mixture was dried over anhydroussodium sulfate and concentrated under vacuum. This resulted in 45 g(92.13%) of tert-butyl (3R)-3-(hydroxymethyl)-4,4-dimethylpentanoate ascolorless oil.

Step 5: Into a 1000-mL 4-necked round-bottom flask purged and maintainedwith an inert atmosphere of nitrogen, was placed tert-butyl(3R)-3-(hydroxymethyl)-4,4-dimethylpentanoate (45.00 g, 208.024 mmol,1.00 equiv), DCM (450.00 mL). This was followed by the addition of TEA(42.10 g, 416.048 mmol, 2.00 equiv) drop wise with stirring at 0° C. in10 min. To this was added MsCl (26.21 g, 228.827 mmol, 1.10 equiv) dropwise with stirring at 0° C. in 10 min. The resulting solution wasstirred for 2 h at 0-10° C. The reaction was then quenched by theaddition of 200 mL of water at 0° C. The resulting solution wasextracted with 2×200 mL of dichloromethane and the organic layerscombined. The resulting mixture was washed with 200 ml of brine. Theresulting mixture was washed with 200 mL of H₂O. The mixture was driedover anhydrous sodium sulfate and concentrated under vacuum. The residuewas applied onto a silica gel column with ethyl acetate/petroleum ether(1:10). The crude product was purified by re-crystallization from PE.This resulted in 25.7709 g (42.08%) of tert-butyl(3R)-3-[(methanesulfonyloxy)methyl]-4,4-dimethylpentanoate as a whitesolid. ESI MS m/z=317.1 [M+Na]⁺. H-NMR: (400 MHz, CDCl₃, ppm): δ4.397-4.383 (1H, m), 4.237-4.215 (1H, m), 3.029 (3H, s), 2.421-2.406(1H, m), 2.319-2.292 (1H, m), 2.165-2.083 (1H, m), 1.482 (9H, s), 0.979(9H, s).

Step 6: A vial was charged with 3,5-dibromo-1H-pyrazole (1 g, 4.56mmol), tert-butyl(3R)-3-[(methanesulfonyloxy)methyl]-4,4-dimethylpentanoate (1 g, 3.43mmol), potassium carbonate (853 mg, 6.18 mmol), and DMSO (10 mL). Themixture was heated to 70° C. for 18 h. After cooling to rt, the mixturewas diluted with water, and the product was extracted with MBTE.Combined organics were concentrated, and the residue was purified onsilica gel to provide tert-butyl(R)-3-((3,5-dibromo-1H-pyrazol-1-yl)methyl)-4,4-dimethylpentanoate (997mg, 69%) as a clear, colorless oil.

Step 7: An oven-dried flask was charged with tert-butyl(R)-3-((3,5-dibromo-1H-pyrazol-1-yl)methyl)-4,4-dimethylpentanoate (997mg, 2.35 mmol) and toluene (30 mL) under an inert nitrogen atmosphere.After cooling to −78° C., a solution of 1.6M nBuLi in hexanes (1.4 mL,2.2 mmol) was added slowly. The reaction mixture was allowed to reachrt, then water was added. The product was extracted with EtOAc, andconcentrated. The residue was purified on silica gel to provide(R)-2-bromo-6-(tert-butyl)-6,7-dihydropyrazolo[1,5-a]pyridin-4(5H)-one(326 mg, 51%).

Step 8: An oven-dried flask under an inert nitrogen atmosphere wascharged with trimethyl orthoformate (1.8 mL, 16.6 mmol) and DCM (10 mL),then cooled to −30° C. Then BF₃—OEt₂ (2.5 mL, 19.9 mmol) was addeddropwise over 15 min. This mixture was allowed to warm to 0° C. over 15min, then cooled to −78° C. Then,(R)-2-bromo-6-(tert-butyl)-6,7-dihydropyrazolo[1,5-a]pyridin-4(5H)-one(900 mg, 3.3 mmol) was added followed by DIPEA (5.8 mL, 33.2 mmol). Thereaction was allowed to reach −10° C. over 90 min. Then, the reactionmixture was poured into sat. NaHCO₃ (250 mL). Then, DCM (150 mL) wasadded. The organic layer was concentrated and the residue was purifiedon silica gel to provide(6R)-2-bromo-6-(tert-butyl)-5-(dimethoxymethyl)-6,7-dihydropyrazolo[1,5-a]pyridin-4(5H)-one(956 mg, 83%).

Step 9: An oven-dried vial under an inert nitrogen atmosphere wascharged with(6R)-2-bromo-6-(tert-butyl)-5-(dimethoxymethyl)-6,7-dihydropyrazolo[1,5-a]pyridin-4(5H)-one(150 mg, 0.4 mmol) and DCM (5 mL). After cooling to −78° C., a solutionof 1M TiCl₄ in DCM (1.3 mL, 1.3 mmol) was added. After 5 min at thistemperature,(Z)-4-methoxy-2,2,8,8-tetramethyl-6-methylene-3,7-dioxa-2,8-disilanon-4-ene(340 mg, 1.3 mmol) was added, then the reaction mixture was allowed toreach rt. After 1 h, MeOH (2 mL) was added and the reaction mixture wasconcentrated and the residue purified on silica gel to provide methyl(S)-2-bromo-6-(tert-butyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylate(120 mg, 14%). ESI MS m/z=380.1 [M+H]⁺.

Step 10: A vial was charged with methyl(S)-2-bromo-6-(tert-butyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylate(35 mg, 0.09 mmol, 1 equiv.), XPhos Pd G3 (8 mg, 0.009 mmol, 0.1equiv.), (4-fluorophenyl)boronic acid (26 mg, 0.185 mmol, 2 equiv.),cesium carbonate (90 mg, 0.28 mmol, 3 equiv.), DMF (5 mL), and water (1mL). The reaction mixture was stirred at 110° C. for 16 h. After coolingto room temperature, the solids were filtered, the filtrate wasconcentrated and purified by RPHPLC to provide(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (11 mg, 31%) as a white solid. ESI MS m/z=381.1 [M+H]⁺.

Ex2: Synthesis of(S)-6-(tert-butyl)-9-hydroxy-2-phenyl-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that phenylboronic acid was used in place of(4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-9-hydroxy-2-phenyl-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (5 mg). ESI MS m/z=363.1 [M+H]⁺.

Ex3: Synthesis of(S)-6-(tert-butyl)-2-(2-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that (2-fluorophenyl)boronic acid was used inplace of (4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-2-(2-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (4 mg). ESI MS m/z=381.1 [M+H]⁺.

Ex4: Synthesis of(S)-6-(tert-butyl)-2-(3-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that (3-fluorophenyl)boronic acid was used inplace of (4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-2-(3-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (7 mg). ESI MS m/z=381.1 [M+H]⁺.

Ex5: Synthesis of(S)-6-(tert-butyl)-1,2-bis(3-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that 3,5-dibromo-4-chloro-1H-pyrazole was usedinstead of 3,5-dibromo-1H-pyrazole in Step 6, and(3-fluorophenyl)boronic acid was used in place of(4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-1,2-bis(3-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (3 mg). ESI MS m/z=475.1 [M+H]⁺.

Ex6: Synthesis of(S)-6-(tert-butyl)-1,2-bis(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that 3,5-dibromo-4-chloro-1H-pyrazole was usedinstead of 3,5-dibromo-1H-pyrazole in Step 6 to provide(S)-6-(tert-butyl)-1,2-bis(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (2 mg). ESI MS m/z=475.1 [M+H]⁺.

Ex7: Synthesis of(S)-6-(tert-butyl)-1,2-bis(2-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that 3,5-dibromo-4-chloro-1H-pyrazole was usedinstead of 3,5-dibromo-1H-pyrazole in Step 6, and(2-fluorophenyl)boronic acid was used in place of(4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-1,2-bis(2-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (4 mg). ESI MS m/z=475.1 [M+H]⁺.

Ex8: Synthesis of(S)-6-(tert-butyl)-9-hydroxy-1,2-diphenyl-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that 3,5-dibromo-4-chloro-1H-pyrazole was usedinstead of 3,5-dibromo-1H-pyrazole in Step 6, and phenylboronic acid wasused in place of (4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-9-hydroxy-1,2-diphenyl-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (5 mg). ESI MS m/z=439.1 [M+H]⁺.

Ex9: Synthesis of(S)-6-(tert-butyl)-1,2-di(furan-3-yl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid

The procedure for the synthesis of the title molecule was the same asfor the synthesis of(S)-6-(tert-butyl)-2-(4-fluorophenyl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (Example 1), except that 3,5-dibromo-4-chloro-1H-pyrazole was usedinstead of 3,5-dibromo-1H-pyrazole in Step 6, and furan-3-boronic acidwas used in place of (4-fluorophenyl)boronic acid in Step 10 to provide(S)-6-(tert-butyl)-1,2-di(furan-3-yl)-9-hydroxy-5,6-dihydropyrazolo[5,1-a]isoquinoline-8-carboxylicacid (6 mg). ESI MS m/z=419.1 [M+H]⁺.

Biological Activity

Methods:

2.2.15 cells are passaged upon attaining confluency in DMEM/F12 media inthe presence of 10% FBS, Penn/Strep, and 250 ug/mL G418. Novel compoundsare 5 fold serially diluted in DMSO and added to 96 well platescontaining 35,000 cells/well at a 1:200 dilution so that the finalconcentration of DMSO is 0.5%. On day 5, post treatment cell lysates andsupernatants are harvested for analysis.

Cells are lysed using Agilent Sidestep Lysis buffer, diluted 1:100 andquantified via quantitative real time PCR. Commercially available ELISAkits are used to quantitate the viral proteins HBsAg (Alpco) or HBeAg(US Biological) by following the manufacturer's recommended protocolafter diluting samples to match the linear range of their respectiveassays. Irrespective of readout, compound concentrations that reduceviral product accumulation in the cell lysates or supernatants by 50%relative to no drug controls (EC₅₀) are reported; EC₅₀ ranges are asfollows: A<0.1 μM; B 0.2-1 μM; C>1 μM.

Additionally, compound induced cellular toxicity is evaluated byexposing HepG2 cells seeded at 5,000 cells/well to serially dilutedcompound with a final DMSO concentration of 0.5% for three days. At day3, post seeding cells are treated with ATPlite 1Step according to themanufacturer's instructions. Compound concentrations that reduce totalATP levels in wells by 50% relative to no drug controls (CC₅₀) arereported; CC₅₀ ranges are as follows: A>25 μM; B 10-25 μM; C<10 μM.

TABLE 3 Summary of Activities Example 2.2.15 cells HepG2 cells NumberEC₅₀ (μM) CC₅₀ (μM) 1 C — 2 C — 3 C — 4 C — 5 C — 6 A A 7 A A 8 A A 9 C—

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed:
 1. A compound represented by Formula (I):

or a pharmaceutically acceptable salt, N-oxide, ester or prodrugthereof, wherein: Q₁, Q₂, Q₃, and Q₄ are each independently selectedfrom hydrogen, halo, —NR₁₁R₁₂, optionally substituted —C₁-C₆ alkyl,optionally substituted —C₂-C₆ alkenyl, optionally substituted —C₁-C₆alkoxy, optionally substituted —C₃-C₈ cycloalkyl; optionally substituted—C₃-C₈ cycloalkenyl; optionally substituted 3- to 8-memberedheterocycloalkyl; optionally substituted aryl; and optionallysubstituted heteroaryl; provided that at least one of Q₃ and Q₄ is nothydrogen; Y₁ is hydrogen, halo, optionally substituted C₁-C₆ alkyl,optionally substituted —C₃-C₆ cycloalkyl, optionally substituted 3- to6-membered heterocycloalkyl, —CN, —C(O)R₁₁; —C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂);—C(O)N(R₁₁)S(O)₂(R₁₂); —S(O)₂R₁₁; or —S(O)₂N(R₁₁)(R₁₂); Y₂ is hydrogen,halo, optionally substituted C₁-C₆ alkyl, optionally substituted —C₃-C₆cycloalkyl, optionally substituted 3- to 6-membered heterocycloalkyl,—CN, —C(O)R₁₁; —C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂); —C(O)N(R₁₁)S(O)₂(R₁₂);—S(O)₂R₁₁; —S(O)₂N(R₁₁)(R₁₂), or —O—Si(R₁₁)₃; Y₃ is —C(O)R₁₁, —COOR₁₁,—C(O)NHSO₂R₁₁, —C(O)NHSO₂NR₁₁R₁₂, or 1,2,4-oxadiazol-3-yl-5(4H)-one, orY₃ is optionally substituted aryl, optionally substituted heteroaryl,optionally substituted —C₅-C₆ cycloalkyl, or optionally substituted 5-to 6-membered heterocycloalkyl; Y₄ is hydrogen, halo, optionallysubstituted C₁-C₆ alkyl, optionally substituted —C₃-C₆ cycloalkyl,optionally substituted 3- to 6-membered heterocycloalkyl, —CN, —C(O)R₁₁;—C(O)OR₁₁; —C(O)N(R₁₁)(R₁₂); —C(O)N(R₁₁)S(O)₂(R₁₂); —OR₁₁, —NR₁₁R₁₂;—SR₁₁; —S(O)₂R₁₁; or —S(O)₂N(R₁₁)(R₁₂); alternatively, Y₂ and Y₃ aretaken together to form an optionally substituted 5-12 memberedheterocyclic ring or carbocyclic ring containing 1, 2, or 3 doublebonds; Z₁ is CR₁, Z₂ is CR₂, and Z₃ is N; R₁ and R₂ are eachindependently selected from: 1) hydrogen; 2) halogen; 3) —NO₂; 4) Cyano;5) Optionally substituted —C₁-C₈ alkyl; 6) optionally substituted —C₂-C₈alkenyl; 7) optionally substituted —C₂-C₈ alkynyl; 8) optionallysubstituted —C₃-C₈ cycloalkyl; 9) optionally substituted 3- to12-membered heterocycloalkyl; 10) optionally substituted aryl; 11)optionally substituted arylalkyl; 12) optionally substituted heteroaryl;13) optionally substituted heteroarylalkyl; 14) —SR₁₁; 15) —S(O)₂R₁₁;16) —S(O)₂N(R₁₁)(R₁₂); 17) —C(O)R₁₁; 18) —C(O)OR₁₁; 19)—C(O)N(R₁₁)(R₁₂); 20) —C(O)N(R₁₁)S(O)₂(R₁₂); 21) —N(R₁₁)(R₁₂); 22)—N(R₁₃)C(O)N(R₁₁)(R₁₂); 23) —N(R₁₁)C(O)(R₁₂); 24) —N(R₁₁)C(O)₂(R₁₂); 25)—N(R₁₃)S(O)₂N(R₁₁)(R₁₂); 26) —N(R₁₁)S(O)₂(R₁₂); 27) —OR₁₁; 28)—OC(O)R₁₁; 29) —OC(O)OR₁₁; and 30) —OC(O)N(R₁₁)(R₁₂); and R₁₁, R₁₂, andR₁₃, are each independently selected from hydrogen, optionallysubstituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl,optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to8-membered heterocycloalkyl, optionally substituted aryl, and optionallysubstituted heteroaryl; alternatively, R₁₁ and R₁₂ are taken togetherwith the nitrogen atom to which they attached to form an optionallysubstituted 3-8 membered heterocyclic containing 0, 1, 2, or 3 doublebonds.
 2. The compound of claim 1, wherein R₁ and R₂ are eachindependently selected from one of the following by removal of ahydrogen atom:

wherein each of these groups is optionally substituted with one or twogroups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substitutedC₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic;and R₁₁ and R₁₂ are as defined in claim
 1. 3. The compound of claim 1,represented by Formula (IV-1) or Formula (IV-2), or a pharmaceuticallyacceptable salt, N-oxide, or ester thereof:

wherein Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, and Z3 are as defined in claim
 1. 4. Acompound of claim 1, represented by Formula (IX-3) or Formula (IX-6), ora pharmaceutically acceptable salt, N-oxide, or ester thereof:

wherein Y₁, Y₂, R₁, and R₂ are as defined in claim
 1. 5. The compound ofclaim 1, represented by one of Formulae (XVIII-4) to (XVIII-6), or apharmaceutically acceptable salt, N-oxide, or ester thereof:

wherein R₂₂ and R₂₃ are each independently selected from halo, CN,—OR₁₁, —NR₁₁R₁₂, and optionally substituted C₁-C₆ alkyl; m is 0, 1, 2, 3or 4; n is 0, 1, 2, 3, or 4; and R₁₁, R₁₂, Y₁, Y₂, Y₃, Y₄, Q₃, and Q₄are as defined in claim
 1. 6. The compound of claim 1, represented byone of Formulae (XX-4) to (XX-6), or a pharmaceutically acceptable salt,N-oxide, or ester thereof:

wherein R₂₂ and R₂₃ are each independently selected from halo, CN,—OR₁₁, —NR₁₁R₁₂, and optionally substituted C₁-C₆ alkyl; m is 0, 1, 2, 3or 4; n is 0, 1, 2, 3, or 4; and R₁₁, R₁₂, Y₁, Q₃, and Q₄ are as definedin claim
 1. 7. The compound of claim 1, selected from the compounds setforth below or a pharmaceutically acceptable salt, N-oxide, or esterthereof: Compound Structure Compound Structure 1

2

3

4

5

6

7

8

9


8. A pharmaceutical composition, comprising a compound according toclaim 1, in combination with a pharmaceutically acceptable carrier orexcipient.
 9. A method of treating an HBV infection in a subject in needthereof, comprising administering to the subject a therapeuticallyeffective amount of a compound or a combination of compounds accordingto claim 1.